U.S. patent application number 10/565079 was filed with the patent office on 2006-10-05 for device for counting cells and method for manufacturing the same.
This patent application is currently assigned to Digital Bio Technology. Invention is credited to Hyun Woo Bang, Jun Keun Chang, Chanil Chung, Seok Chung, Han Sang Jo.
Application Number | 20060223165 10/565079 |
Document ID | / |
Family ID | 36242023 |
Filed Date | 2006-10-05 |
United States Patent
Application |
20060223165 |
Kind Code |
A1 |
Chang; Jun Keun ; et
al. |
October 5, 2006 |
Device for counting cells and method for manufacturing the same
Abstract
Disclosed are a device for counting cells and a method for
manufacturing the device. The device comprises a transparent upper
substrate and a transparent lower substrate. Fine lattice patterns
for counting cells are formed on the lower substrate. A space for
filling the sample, including cells, is provided between the upper
substrate and the lower substrate. A hole for injecting the samples
into the space for filling the samples is provided in the upper
substrate. The device is disposable, and the cost of manufacturing
the device is low. It is easy to fill the samples into the filling
space and to count cells by using the device. The upper substrate
and the lower substrate are bonded, and thus, no cover is
required.
Inventors: |
Chang; Jun Keun; (Seoul,
KR) ; Chung; Seok; (Seoul, KR) ; Chung;
Chanil; (Kyunggi-Do, KR) ; Bang; Hyun Woo;
(Seoul, KR) ; Jo; Han Sang; (Kyunggi-Do,
KR) |
Correspondence
Address: |
BROWDY AND NEIMARK, P.L.L.C.;624 NINTH STREET, NW
SUITE 300
WASHINGTON
DC
20001-5303
US
|
Assignee: |
Digital Bio Technology
Seoul National University Institute of Advanced, Machinery,
1304, San 56-1 Shinlim, Kawan
Seoul
KR
151-742
|
Family ID: |
36242023 |
Appl. No.: |
10/565079 |
Filed: |
July 13, 2004 |
PCT Filed: |
July 13, 2004 |
PCT NO: |
PCT/KR04/01735 |
371 Date: |
January 18, 2006 |
Current U.S.
Class: |
435/287.1 ;
156/219 |
Current CPC
Class: |
B82Y 15/00 20130101;
G01N 33/49 20130101; Y10T 156/1039 20150115; G01N 15/1456 20130101;
G02B 21/34 20130101; G01N 2015/1486 20130101 |
Class at
Publication: |
435/287.1 ;
156/219 |
International
Class: |
C12M 1/34 20060101
C12M001/34; B44C 3/08 20060101 B44C003/08 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 18, 2003 |
KR |
10 2003 0049406 |
Claims
1. A device for counting fine particles comprising, a transparent
lower substrate having fine lattice patterns for counting the fine
particles formed on an upper surface thereof; and a transparent
upper substrate stacked on the lower substrate, wherein the upper
substrate comprises a fill chamber having a predetermined height
from a bottom surface of the upper substrate and forming a space
for filling a sample including the fine particles on the fine
lattice patterns and an injecting hole for the sample communicated
with the fill chamber.
2. The device according to claim 1, wherein the upper substrate
further comprises a discharge hole communicated with the fill
chamber for discharging the sample or an air bubble from the fill
chamber.
3. The device according to claim 1, wherein the upper and lower
substrates are bonded and thus form an integrated body.
4. The device according to claim 3, wherein the upper and lower
substrates are bonded by a heating, an adhesive, a coating, a
pressurization, a vibration or an ultrasonic bonding.
5. The device according to claim 1, wherein the fill chamber is
formed with a height of 50-200 .mu.m.
6. The device according to claim 1, wherein an area of the fill
chamber in the upper and lower substrates is transparent and the
fine lattice patterns are formed in a predetermined place of the
area in which the fill chamber is formed on the lower
substrate.
7. The device according to claim 1, wherein an indicative member is
formed on the upper substrate for indicating a position of the fine
lattice patterns.
8. The device according to claim 1, wherein the upper or lower
substrate is made of plastics.
9. The device according to claim 1, wherein the fine particles are
blood cells or bacteria.
10. A manufacturing method of a device for counting fine particles
comprising steps of; forming fine lattice patterns on a
predetermined place of a lower substrate; forming a fill chamber
having a predetermined height for filling a sample including the
fine particles, an injecting hole and a discharge hole communicated
with the fill chamber in an upper substrate; and bonding the upper
and lower substrates.
11. The method according to claim 10, wherein the step of forming
fine lattice patterns on a predetermined place of the lower
substrate comprising steps of: forming a photoresist layer on a
plate; forming a mask pattern having fine lattice patterns on the
plate by patterning the photoresist layer; etching the plate by
using the mask pattern as an etching mask; removing the mask
pattern to obtain the plate as a mold wherein the fine lattice
patterns are formed; pouring melted plastics on the mold, and then
cooling and curing the plastics on the mold; and removing the mold
to obtain the plastics as the lower substrate wherein the fine
lattice patterns are formed.
12. The method according to claim 10, wherein the step of forming
fine lattice patterns on a predetermined place of the lower
substrate comprising steps of: forming a photoresist layer on a
plate; forming a mask pattern having fine lattice patterns on the
plate by patterning the photoresist layer; etching the plate by
using the mask pattern as an etching mask; removing the mask
pattern to obtain the plate as a master wherein the fine lattice
patterns are formed; forming Ni layer on the master by electroless
plating or electrolysis plating; removing the master to obtain the
Ni layer as a mold wherein the fine lattice patterns are formed;
pouring melted plastics on the mold, and then cooling and curing
the plastics on the mold; and removing the mold to obtain the
plastics as the lower substrate wherein fine lattice patterns are
formed.
Description
TECHNICAL FIELD
[0001] The present invention relates to a device for counting
cells, and in particular to a device for counting cells comprising
a transparent lower substrate having fine lattice patterns for
counting the cells formed on an upper surface thereof and a
transparent upper substrate stacked on the lower substrate, wherein
the upper substrate comprises a fill chamber having a predetermined
height from a bottom surface of the upper substrate and forming a
space for filling a sample including the cells into the fine
lattice patterns and an injecting hole for the sample communicated
with the fill chamber.
BACKGROUND ART
[0002] Generally, when diagnosing a disease, it is examined the
number and the functions of typical blood cells such as
erythrocytes, leukocytes or platelets included in the blood. For
example, it is possible to diagnose tuberculosis, obesity or
pregnancy from a blood sedimentation rate and dehydration or anemia
from a corpuscular volume. Also, it is possible to diagnose chronic
leukemia from the number of platelets, kidney disease, hypoxia,
smoking, pulmonary disease, hemolytic anemia or aplastic anemia
from the number of erythrocytes, and acute typhlitis, leukemia or
aplastic anemia from the number of leukocytes. Like this, the
measurement of the number of blood cells is closely related to the
disease diagnosis.
[0003] The size of erythrocytes, which are typical blood cells, is
classified into micro, normal, macro and mega. By finding out the
sizes and the number of erythrocytes, it is possible to use them as
diagnostic materials for various diseases as described above.
[0004] Particularly, it is required to know the number of
erythrocytes for seeking whether or not anemia and cause thereof.
For the healthy public, a male has about 4,400,000.about.5,600,000
erythrocytes/dl in blood and a female has about
3,500,000.about.5,000,000 erythrocytes/dl.
[0005] From the measurement of the number of erythrocytes, when the
number of erythrocytes is increased beyond a reference value, it is
possible to diagnose diseases such as polycythemia vera,
dehydration, shock adrenal insufficiency or cardiopulmonary
disease. Also, when the number of erythrocytes is decreased, it is
possible to diagnose whether or not various anemia.
[0006] FIG. 1 is a perspective view showing a device for measuring
the number of blood cells such as erythrocytes according to the
prior art.
[0007] As shown in FIG. 1, the device 10 for measuring the number
of erythrocytes according to the prior art comprises a body 15 made
of glass or quartz, partition walls 20, 25 provided on an upper
part of the body 15, a measurement part 30 formed between the
partition walls 20, 25 and a cover 35 covering an upper part of the
measurement part 30.
[0008] The partition walls 20, 25 located on the body 15 and the
measurement part 30 located between the partition walls 20, 25 are
formed on the body 15 by micromachining the body 15 made of glass
or quartz as a method disclosed in Korean Unexamined Patent
Publication No. 1999-84670, for example.
[0009] The partition walls 20, 25 are upwardly protruded from the
upper part of the body 15 at the periphery of the measurement part
30 so that a sample such as blood is not flown out of the
measurement part 30 when the sample is poured into the measurement
part 30. Also, the cover 35 made of glass is provided on the
partition walls 20,25, so that the sample is in existence in the
measurement part 30 between the partition walls 20, 25 and the
cover 35 and thus the number of cells in the sample, such as blood
cells in the blood, is measured.
[0010] FIG. 2 is a schematic plan view showing the measurement part
of the device shown in FIG. 1.
[0011] As shown in FIGS. 1 and 2, the measurement part 30 consists
of a plurality of measuring areas 45 and bright lines 40 for
distinguishing each of the measuring areas 45.
[0012] Generally, the bright lines 40 are arranged in a cross
pattern and thus divide the measurement part 30 into four measuring
areas 45. A plurality of lattice lines, which are arranged
lengthwise and crosswise according to the sizes of cells to be
measured, are formed in each measuring area 45. For example, blood
is dropped in the measuring areas 45 having such a construction and
thus the number of cells, which are in existence between the
lattice lines, is measured.
[0013] However, in the device for measuring the number of cells
such as erythrocytes, since the body made of glass or quartz is
relatively expensive and it takes much time and efforts to
micromachine the body, the time and cost for manufacturing the
device are increased. Like this, since the prior device for
measuring the number of cells is expensive, it is required that
once the device is used, it should be washed and then reused.
Accordingly, it should be put up with inconveniences of washing the
device and there is a possibility that the sample previously
measured remains in the device.
[0014] Also, since the device made of glass or quartz is fragile by
an impact, there is some danger that the device is damaged during
using it.
[0015] In addition, since the body 15 and the cover 35 are
separated, there is troublesomeness that the sample should be
dropped after covering the body with the cover. Particularly, since
the tightness between the cover and the partition walls is very
poor, it is required that a very strong force should be applied to
the cover or the cover should be attached to the partition walls
with a separate adhesive. Also, there is a possibility that the
cover is damaged during using the device even if such an additional
process was used.
DISCLOSURE OF INVENTION
[0016] Accordingly, the present invention has been made to solve
the above-mentioned problems occurring in the prior art. The object
of the present invention is to provide a device for counting cells.
The other object of the invention is to provide a manufacturing
method of the device.
[0017] In order to accomplish the objects, the invention comprises
a transparent lower substrate having fine lattice patterns for
counting cells formed on an upper surface thereof; and a
transparent upper substrate stacked on the lower substrate, wherein
the upper substrate comprises a fill chamber having a predetermined
height from a bottom surface of the upper substrate and forming a
space for filling a sample including the cells on the fine lattice
patterns and an injecting hole for the sample communicated with the
fill chamber.
[0018] When the upper substrate and the lower substrate are
integratedly made by bonding them, a covering process as the prior
device for counting cells is not required. Accordingly, since it is
easy to fill the fill chamber with the sample by dropping the
sample into the injection hole, the device can be more easily used
than that of the prior art. Also, since the cost of manufacturing
the device for counting cells can be greatly decreased, the device
can be disposably and easily used.
[0019] The invention relates to a device for counting fine
particles such as cells.
[0020] More specifically, the invention provides a device for
counting fine particles comprising a transparent lower substrate
having fine lattice patterns for counting the fine particles formed
on an upper surface thereof; and a transparent upper substrate
stacked on the lower substrate, wherein the upper substrate
comprises a fill chamber having a predetermined height from a
bottom surface of the upper substrate and forming a space for
filling a sample including the fine particles on the fine lattice
patterns and an injecting hole for the sample communicated with the
fill chamber.
[0021] Preferably, the upper substrate further comprises a
discharge hole communicated with the fill chamber for discharging
the sample or an air bubble from the fill chamber.
[0022] Particularly, the upper and lower substrates are preferably
bonded and thus form an integrated body. The upper and lower
substrates are bonded by a convenient method such as a heating, an
adhesive, a coating, a pressurization or a vibration, preferably an
ultrasonic bonding.
[0023] According to the invention, a height of the fill chamber may
be arbitrarily formed according to a volume of the sample to be
examined. The height is preferably 50-200 .mu.m and most preferably
100 .mu.m.
[0024] According to the invention, an area of the fill chamber in
the upper and lower substrates is made to be transparent for a
microscopic observation. The fine lattice patterns are formed in a
predetermined place of the area in which the fill chamber is formed
on the lower substrate. It is possible to calculate a volume of the
fill chamber by making the area of the fill chamber have a
predetermined area and a predetermined height.
[0025] An indicative member is preferably formed on the upper
substrate for indicating a position of the fine lattice patterns.
Accordingly, when counting the cells in the sample with a
microscope, it is possible to easily find the position of the fine
lattice patterns.
[0026] The upper substrate or lower substrate may be made by an
arbitrary material, referably any plastics capable of being
injection-molded such as polycarbonate (PC), olymethylmethacrylate
(PMMA), polyethylene (PE), polyethyleneterephthalate PET) or
polystyrol (PS).
[0027] By using the device according to the invention, it is
possible to easily count rythrocytes, leukocytes or platelets
included in the blood. Additionally, it is possible o easily count
microbes which are unicellular organisms, bacteria and any fine
articles.
[0028] In addition, the invention provides a manufacturing method
of a device for counting fine particles comprising steps of forming
fine lattice patterns on a redetermined place of a lower substrate;
forming a fill chamber having a predetermined height for filling a
sample including the fine particles, such as blood cells or
bacteria, an injecting hole and a discharge hole communicated with
the fill chamber in an upper substrate; and bonding the upper and
lower substrates.
BRIEF DESCRIPTION OF DRAWINGS
[0029] The above and other objects, features and advantages of the
present invention will be more apparent from the following detailed
description taken in conjunction with the accompanying drawings, in
which:
[0030] FIG. 1 is a perspective view showing a device for measuring
the number of erythrocytes according to the prior art;
[0031] FIG. 2 is a schematic plan view showing a measurement part
of the device shown in FIG. 1;
[0032] FIG. 3 is a perspective view of an upper substrate of a
device according to an embodiment of the invention;
[0033] FIG. 4 is a sectional view of the upper substrate shown in
FIG. 3;
[0034] FIG. 5 is a plan view of the upper substrate shown in FIG.
3;
[0035] FIG. 6 is a perspective view of a lower substrate of the
device according to an embodiment of the invention;
[0036] FIG. 7 shows fine lattice patterns formed on the lower
substrate;
[0037] FIG. 8 shows an embodiment of the invention, in which upper
and lower substrates are bonded;
[0038] FIG. 9 shows another embodiment of the invention;
[0039] FIG. 10a to 10d are sectional views for illustrating an
example of a process of forming fine lattice patterns on the lower
substrate;
[0040] FIG. 11a to 11h are sectional views for illustrating another
example of a process of forming fine lattice patterns on the lower
substrate.
DESCRIPTION OF REFERENCE NUMERALS FOR IMPORTANT PART OF THE
DRAWINGS
[0041] TABLE-US-00001 100: upper substrate 110: fill chamber 120:
injecting hole 130: discharge hole 140: indicative member 200:
lower substrate 210: fine lattice patterns
BEST MODE FOR CARRYING OUT THE INVENTION
[0042] Hereinafter, preferred embodiments of the present invention
will be described with reference to the accompanying drawings. In
the following description of the present invention, a detailed
description of known functions and configurations incorporated
herein will be omitted when it may make the subject matter of the
present invention rather unclear.
[0043] FIG. 3 is a perspective view showing an upper substrate of a
device according to an embodiment of the invention, FIG. 4 is a
sectional view of the upper substrate and FIG. 5 is a plan view of
the upper substrate.
[0044] As shown in FIGS. 3 to 5, the upper substrate 100 comprises
a fill chamber 110 having a predetermined height from a bottom
surface of the upper substrate and forming a space for filling a
sample, an injecting hole 120 for the sample communicated with the
fill chamber and a discharge hole 130 for discharging the air and
the excess sample in the fill chamber 110 when injecting the
sample. In addition, an indicative member 140 is formed on the
upper substrate 100 for indicating a position of fine lattice
patterns formed on a lower substrate.
[0045] The sample can be easily injected when the injecting hole
120 and the discharge hole 130 are provided at opposite
positions.
[0046] The upper substrate is made of transparent plastics and can
be manufactured by a typical injection molding.
[0047] FIG. 6 is a perspective view showing a lower substrate of
the device according to an embodiment of the invention. Transparent
plastics capable of being injection-molded is used for the lower
substrate.
[0048] Fine lattice patterns 210 for counting cells of the sample
are formed on the lower substrate. The upper substrate is stacked
on the lower substrate, so that the fill chamber 110 is formed. A
method for forming the fine lattice patterns will be described
later.
[0049] FIG. 7 is an enlarged view of the fine lattice patterns
formed on the lower substrate. Shape, height, width and interval,
etc. of the fine lattice patterns can be arbitrarily formed as
necessary. Preferably, the height, the width and the interval of
the fine lattice patterns are about 1 .mu.m, about 1.5 .mu.m and 10
.mu.m, respectively.
[0050] FIG. 8 shows a device according to an embodiment of the
invention, which is integratedly made by bonding the upper and
lower substrates with an ultrasonic bonding.
[0051] FIG. 9 shows a device according to another embodiment of the
invention, which device comprises two fill chambers 111, 112
separated by a partition wall. Thus, injecting holes 121, 122,
discharge holes 131, 132 and indicative members 141, 142 are
separately formed in each of the fill chambers 111, 112. As shown
in FIG. 9, the device may comprise at least two fill chambers as
necessary.
[0052] Hereinafter, a method for forming fine lattice patterns on
the lower substrate will be described.
[0053] FIGS. 10a to 10d show an example of a process for forming
fine lattice patterns on the lower substrate.
[0054] Firstly, as shown in FIG. 10a, a plate 310 made of glass,
silicon or ceramics is provided. A layer 320 of photoresist is
formed on the plate, for example, by a spin coating. The plate 310
is used as a mold for molding a lower substrate.
[0055] Then, the layer of photoresist is patterned by exposure and
developing processes, so that a mask pattern 320 having fine
lattice patterns is formed on the plate.
[0056] Then, the plate 310 is etched by using the mask pattern 320
as an etching mask and removed by a strip process, resulting in a
mold 310 having fine lattice patterns formed as shown in FIG.
10b.
[0057] As shown in FIG. 10c, melted state of plastics 200 which is
heated to a predetermined temperature is poured in the mold 310.
Then, the melted plastics 200 is cooled and cured in the mold
310.
[0058] After the plastics 200 is cured in the mold 310, the mold
310 is separated from the cured plastics 200, so that a lower
substrate 200 having fine lattice patterns 210 formed is
manufactured, as shown in FIG. 10d.
[0059] With the above method, the plate 310 itself is used as a
mold. However, after an oxide, nitride or metal layer is
additionally deposited on the plate 310 as a mold-forming layer,
the mold-forming layer is formed with fine lattice patterns and can
then be used as a mold.
[0060] FIGS. 11a to 11h show sectional views for illustrating
another example of a process forming fine lattice patterns on the
lower substrate. In this example, contrary to the process shown in
FIGS. 10a to 10d, a master for forming a mold is separately
manufactured.
[0061] Firstly, as shown in FIG. 11a, a layer 420 of photoresist is
formed on a plate 410 of glass, silicon or ceramics, which is used
as a master, by a spin coating method, for example.
[0062] After that, as shown in FIG. 11b, the layer 420 of
photoresist is patterned by exposure and developing processes.
[0063] Then, as shown in FIG. 11c, the plate 410 is etched by using
the patterns 420 of the photoresist as an etching mask.
[0064] After that, as shown in FIG. 11d, the mask 420 is removed by
a strip process, so that a master 410 having fine lattice patterns
formed is provided.
[0065] Then, as shown in FIG. 11e, a Ni-layer 430 is formed on the
master 410 by an electroless plating or electrolysis plating
method. After that, the master 410 is removed, so that a mold 430
made of Ni is provided as shown in FIG. 11f. At this time, just
before the plating step, the master is preferably surface-treated
by a sputtering, vacuum vapor-deposition or non-electrolytic
plating process so that the master 410 is electrically
conducted.
[0066] Then, as shown in FIGS. 11g and 11h, a lower substrate 200
having fine lattice patterns 210 formed can be manufactured by a
molding process using the mold 430.
[0067] Preferably, the upper or lower substrate made as described
above is passed through an additional process such as a hydrophilic
treatment or reactive group introduction. When the device of the
invention is treated with oxygen-plasma, etc. to make the device
hydrophilic, aqueous liquid such as blood can flow well and
uniformly spread on the surface thereof. In addition, in order to
introduce desired reactive group, for example, amine group, the
device can be treated with plasma of the amine group or other
chemical method (surface modification). Like this, when the device
according to the invention is surface-treated, its performance is
further improved.
INDUSTRIAL APPLICABILITY
[0068] As described above, since the device of the invention is
integratedly made by bonding the upper and lower substrates, a
covering process as the prior device for counting cells is not
required. Accordingly, since it is easy to fill the fill chamber
with the sample by dropping the sample into the injection hole, the
device can be more easily used than that of the prior art. Also,
since the cost of manufacturing the device for counting cells is
greatly decreased, the device can be disposably and easily
used.
[0069] While the invention has been shown and described with
reference to certain preferred embodiments thereof, it will be
understood by those skilled in the art that various changes in form
and details may be made therein without departing from the spirit
and scope of the invention as defined by the appended claims.
* * * * *